OVERALL SUMMARY This U54 ?Immuno-engineering to Improve Immunotherapy (i3) Center? was developed in response to RFA-CA-19-013. The purpose of this U54 is to employ immuno-engineering principles to design more durable, accessible, and less toxic immunoprevention and immunotherapy strategies. A national priority set by the Cancer Moonshot initiative is to support studies incorporating next-generation genetic engineering, engineered biomaterials, nanotechnology, computational and mathematical modelling, or systems biology approaches in the design, implementation and evaluation of next generation immunotherapies focused on overcoming immunosuppressive barriers, targeting tumor cells for killing, and generating long-lasting anti-cancer immunological memory. The focus of our U54 entitled ?Engineering the next generation of T cells? is on developing next-generation gene-editing or modification of immune cells to improve persistence in vivo, control and manipulate the immune system to reduce o?-target toxicities and enhance anti-tumor e?ectiveness of adoptive cell therapy. Our central hypothesis is that next generation engineering can improve the safety and efficacy of CAR T cells while decreasing the cost of goods. There are three scientific projects in our U54 Center. The shared goal of these projects is to enable this powerful therapy to reach a broader spectrum of patients with blood cancer and solid cancers. In Project 1, we are using CRISPR/Cas9 to edit the epigenome of T cells and to make cells resistant to cytokine release syndrome (CRS), while employing engineering technologies to automate CAR T cell culture. The goal of Project 2 is to generate universal CAR T cells (UCART) using advanced genetic editing technologies and then to compare UCAR to autologous CAR T in a unique model of canine cancer. In Project 3 we will use advanced protein engineering, cell engineering and oncolytic vectors to test synthetic cytokine and cytokine receptor systems in mouse and human T cells, with the long-term goal of eliminating or reducing the need for lymphodepleting conditioning chemotherapy prior to adoptive cell transfer. Our approach is multidisciplinary and multi- institutional. We have brought together a cadre of exceptional investigators from multiple disciplines who have collaborated and published together for many years. State of the art genetic editing with CRISPR/Cas9 and cell culture technologies at the University of Pennsylvania are synergistically coupled with world-class protein engineering and structural biology at Stanford University. Each project is led by recognized authorities in the field. The Administrative Core for this U54 is designed to manage and coordinate all i3 Center Research Project activities and serve as the liaison between the i3 Center and the IOTN Data Management and Resource-sharing Center (U24) and other Cancer Moonshot consortia, including the Pediatric Immunotherapy Discovery and Development Network (PI-DDN). In addition, the Administrative Core will coordinate collaborative research activities between the i3 Center Research Project PD(s)/PI(s) and IOTN (or other) investigators. The potential for paradigm-shifting impact this U54 is to transform the lessons of first-generation CAR T for leukemia and lymphoma into meaningful efficacy against all malignancies.